文献类型：期刊文献

中文题名：鸡传染性法氏囊病病毒VP3蛋白的原核表达及其B细胞抗原表位鉴定

英文题名：Identification of B cell epitopes of infectious bursal disease virus VP3 expressed in E.coli

作者：赵坤[1];郑玉姝[1];赵德明[2];赵朴[1];赵恒章[1];常新耀[1];王选年[1,3,4]

第一作者：赵坤

机构：[1]河南科技学院动物科学学院;[2]中国农业大学动物医学院;[3]新乡学院;[4]河南省农科院河南省动物免疫学重点实验室

第一机构：河南科技学院动物科学学院,河南新乡453003

年份：2009

期号：12

起止页码：958-962

中文期刊名：中国预防兽医学报

外文期刊名：Chinese Journal of Preventive Veterinary Medicine

收录：CSTPCD;;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

基金：河南省高校杰出科研人才创新工程(2005KYCX008)

语种：中文

中文关键词：鸡传染性法氏囊病病毒;VP3蛋白;原核表达;重叠多肽;B细胞抗原表位

外文关键词：infectious bursal disease virus; VP3 protein; prokaryotic expression; overlapping peptides; B cell epitope

摘要：为鉴定鸡传染性法氏囊病病毒(IBDV)VP3蛋白中的B细胞抗原表位,本研究将IBDV的VP3基因亚克隆于pET-28a中,构建了表达重组质粒pETVP3,经IPTG诱导在E. coli BL21(DE3)中表达了重组蛋白(rVP3)。Western blot鉴定表明,rVP3能被IBDV抗血清特异性识别。同时,根据IBDVVP3的氨基酸序列,合成覆盖VP3全序列的重叠多肽,并与载体蛋白BSA藕联制备多肽人工结合抗原。Peptide-ELISA和Dot-ELISA检测结果表明VP3中有2个线性表位可以被已制备的单克隆抗体(MAb)识别,即728PRDWDRLPYLNL739和982PKPKPKPNAPTQ993;Dot-ELISA结果显示,在VP3中还存在另外4个线性多克隆抗体识别位点:818LANAPQAGSKSQRA831,851QREKD TRISKKMETMGIYFATP872,876ALNGHRGPSPGQLKYWQNTREI897和961QMKDLLLTAMEMK973。这些抗原表位的鉴定为开发IBD表位疫苗奠定了基础。
To identificate B cell epitopes on the VP3 protein of infectious bursal disease vires （IBDV）, the VP3 gene of IBDV was subcloned into the pET28a vector and expressed in E. coil BL21 （DE3） by inducing with IPTG. The expressed VP3 protein could be recognized specifically by anti-IBDV antisera in western blot assay. Overlapping peptides that span the full sequence of VP3 were synthesized and conjugated to carrier protein BSA to prepare conjugated antigens. Their reactivity with anti-IBDV monoclonal antibodies （MAb） or polyclonal antibody was detected by peptide-ELISA and Dot-ELISA. Two linear epitopes, ^728PRDWDRLPYLNL^739 and ^982PKPKPKPNAPTQ^993, were identified by VP3 specific neutralizing MAbs, which could also react with IBDV antiserum. Dot-ELISA confirmed that there were 4 more linear epitopes recognized by polyclonal antibody against the VP3 protein. These epitopes could be developed as potential IBD epitope vaccine.